A Novel Nonsense Mutation Associated with an Exon Skipping in a Patient with Hereditary Protein S Deficiency Type I

Yoshihiro Okamoto; Tomio Yamazaki; Akira Katsumi; Tetsuhito Kojima; Junki Takamatsu; Mikio Nishida; Hidehiko Saito

doi:10.1055/s-0038-1650387

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 1996; 75(06): 877-882
DOI: 10.1055/s-0038-1650387

Original Article

Schattauer GmbH Stuttgart

A Novel Nonsense Mutation Associated with an Exon Skipping in a Patient with Hereditary Protein S Deficiency Type I

Yoshihiro Okamoto

¹The First Department of Internal Medicine, Nagoya University School of Medicine, Nagoya, Japan

³Department of Clinical Pharmacy, Faculty of Pharmacy, Meijo University, Nagoya, Japan

,

Tomio Yamazaki

¹The First Department of Internal Medicine, Nagoya University School of Medicine, Nagoya, Japan

,

Akira Katsumi

¹The First Department of Internal Medicine, Nagoya University School of Medicine, Nagoya, Japan

,

Tetsuhito Kojima

¹The First Department of Internal Medicine, Nagoya University School of Medicine, Nagoya, Japan

,

Junki Takamatsu

²Department of Transfusion Medicine, Nagoya University Hospital, Nagoya, Japan

,

Mikio Nishida

³Department of Clinical Pharmacy, Faculty of Pharmacy, Meijo University, Nagoya, Japan

,

Hidehiko Saito

¹The First Department of Internal Medicine, Nagoya University School of Medicine, Nagoya, Japan

› Author Affiliations

Abstract

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Summary

The genetic defect in a patient with hereditary type I protein S (PS) deficiency was investigated. All the exons and intron-exon junctions of the patient’s PS gene were amplified by PCR and subjected to heteroduplex screening. Only the PCR product of exon 4 revealed heteroduplex bands. A novel nonsense mutation, Ser62 (TCA) to Stop (TGA) was found in exon 4. RT-PCR detected the aberrant mRNA in the patient’s platelets, which was markedly reduced in amount and lacked the region of exon 4, suggesting that the nonsense mutation affected the mutated mRNA metabolism and induced exon skipping. The skipping of exon 4 causes an in-frame deletion of 29 amino acids which just construct the thrombin-sensitive region of the PS molecule. The loss of such an important domain as well as the quantitative decrease in the mutated mRNA appear to be responsible for the type I PS deficiency in this patient.

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References

References
1 Dahlbäck B. Protein S and C4b-binding protein: components involved in the regulation of the protein C anticoagulant system. Thromb Haemost 1991; 66: 49-61
2 Heeb MJ, Mesters RM, Tans G, Rosing J, Griffin JH. Binding of protein S to factor Va associated with inhibition of prothrombinase that is independent of activated protein. C. J Biol Chem 1993; 268: 2872-2877
3 Heeb MJ, Rosing J, Bakker HM, Fernández JA, Tans G, Griffin JH. Protein S binds to and inhibits factor Xa. Proc Natl Acad Sci USA 1994; 91: 2728-2732
4 Comp PC, Esmon CT. Recurrent venous thromboembolism in patients with a partial deficiency of protein. S. N Engl J Med 1984; 311: 1525-1528
5 Comp PC, Nixon RR, Cooper MR, Esmon CT. Familial protein S deficiency is associated with recurrent thrombosis. J Clin Invest 1984; 74: 2082-2088
6 Kamiya T, Sugihara T, Ogata K, Saito H, Suzuki K, Nishioka J, Hashimoto S, Yamagata K. Inherited deficiency of protein S in a Japanese family with recurrent venous thrombosis: a study of three generations. Blood 1986; 67: 406-410
7 Dählback B, Stenflo J. High molecular weight complex in human plasma between vitamin K-dependent protein S and complement component C4b-binding protein. Proc Natl Acad Sci USA 1981; 78: 2512-2516
8 Griffin JH, Gruber A, Fernández JA. Reevaluation of total, free and bound protein S and C4b-binding protein levels in plasma anticoaglated with citrate or hirudin. Blood 1992; 79: 3203-3211
9 Schmidel DK, Tatro AV, Phelps LG, Tomczak JA, Long GL. Organization of the human protein S genes. Biochemistry 1990; 29: 7845-7852
10 Ploos van Amstel HK, Huisman MV, Reitsma PH, ten Cate JW, Bertina RM. Partial protein S gene deletion in a family with hereditary thrombophilia. Blood 1989; 73: 479-483
11 Schmidel DK, Nelson RM, Broxson EH, Comp PC, Marlar RA, Long GL. A 5.3-kb deletion including exon XIII of the protein S α gene occurs in two protein S-deficient families. Blood 1991; 77: 551-559
12 Bertina RM, Pools van Amstel HK, Van Wijngaarden A, Cohnen J, Leem-huis MP, Deutz-Terlouw PP, Van der Linden IK, Reitsma PH. Heerlen polymorphism of protein S, an immunologic polymorphism due to dimorphism of residue 460. Blood 1990; 76: 538-548
13 Diepstraten CM, Ploos van Amstel JK, Reitsma PH, Bertina RM. A CCA/CCG neutral dimorphism in the codon for Pro 626 of the human protein S gene a (PROS1). Nucl Acid Res 1991; 19: 5091
14 Yamazaki T, Sugiura I, Matsushita T, Kojima T, Kagami K, Takamatsu J, Saito H. A phenotypically neutral dimorphism of protein S: the substitution of Lysl55 by Glu in the second EGF domain predicted by an A to G base exchange in the gene. Thromb Res 1993; 70: 395-403
15 Hayashi T, Nishioka J, Shigekiyo T, Saito S, Suzuki K. Protein S Tokushima: abnormal molecule with a substitution of Glu for Lys-155 in the secomd epidermal growth factor-like domain of protein S. Blood 1994; 83: 683-690
16 Reitsma PH, Ploos van Amstel HK, Bertina RM. Three novel mutations in five unrelated subjects with hereditary protein S deficiency type I. J Clin Invest 1994; 93: 486-492
17 Borgel D, Grandrille S, Gouault-Heilmann M, Aiach M. First frameshift mutation in the active protein S gene associated with a quantitative hereditary deficiency. Blood Coagul Fibrinolysis 1994; 5: 593-600
18 Gómez E, Ledford MR, Pegelow CH, Reitsma PH, Bertina RM. Homozygous protein S deficiency due to a one base pair deletion that leads to a stop codon in Exon III of the protein S gene. Thromb Haemost 1994; 71: 723-726
19 Gandrille S, Borgel D, Eschwege-Gufflet V, Aillaud Mf, Dreyfus M, Matheron C, Gaussem P, Abgrall JF, Jude B, Sie P, Toulon P, Aiach M. Identification of 15 different candidate causual point mutations and three polymorphisms in 19 patients with protein S deficiency using a scanning method for the analysis of the protein S active gene. Blood 1995; 85: 130-138
20 Gómez E, Poort SR, Bertina RM, Reitsma PH. Identification of eight point mutations in protein S deficiency type I: analysis of 15 pedigrees. Thromb Haemost 1995; 73: 750-755
21 Yamazaki T, Hamaguchi M, Katsumi A, Kagami K, Kojima T, Takamatsu J, Saito H. A quantitative protein S deficiency associated with a novel nonsense mutation and markedly reduced levels of mutated mRNA. Thromb Haemost 1995; 74: 590-595
22 Formstone CJ, Wacey AI, Berg LP, Rahman S, Bevan D, Rowley M, Voke J, Bemardi F, Legnani C, Simioni P, Girolami A, Tuddenham EGD, Kakkar VV, Cooper DN. Detection and characterization of seven novel protein S (PROS) gene lesions: evaluation of reverse transcript-polymerase chain reaction as a mutation screening strategy. Blood 1995; 86: 2632-2641
23 Dahlbäck B, Carlsson M, Svensson PJ. Familial thrombophilia due to a previously unrecognized mechanism characterized by poor anticoagulant response to activated protein C: prediction of a cofactor to activated protein C. Proc Natl Acad Sci USA 1993; 90: 1004-1008
24 Sugiura I, Matsushita T, Tanimoto M, Takahashi I, Yamazaki T, Yamamoto K, Takamatsu J, Kamiya T, Saito H. Three distinct candidate point mutations of the von Willebrand factor gene in four patients with type IIA von Willebrand disease. Thromb Haemost 1992; 67: 612-617
25 Chowdhury V, Olds RJ, Lane DA, Conard J, Pabinger I, Ryan K, Bauer KA, Bhavnani M, Abildgaard U, Finazzi G, Castaman G, Mannucci PM, Thein SL. Identification of nine novel mutations in type I antithrombin deficiency by heteroduplex screening. Br J Haematol 1993; 84: 656-661
26 Hall GW, Thein S. Nonsense codon mutations in the terminal exon of the β-globin gene are not associated with a reduction in β-mRNA accumulation: a mechanism for the phenotype of dominant b-thalassemia. Blood 1994; 83: 2031-2037
27 Kunishima S, Miura H, Fukutani H, Yoshida H, Osumi K, Kobayashi S, Ohno R, Naoe T. Bemard-Soulier syndorome Kagoshima: Ser 444 → stop mutation of glycoprotein (GP) Iba resulting in circulating truncated GPIba and surface expression of GPIbB and GPIX. Blood 1994; 84: 3356-3362
28 Cooper DN. Human gene mutations affecting RNA processing and translation. Ann Med 1993; 25: 11-17
29 Gibson RA, Hajianpour A, Murer-Oriando M, Buchwald M, Mathew CG. A nonsense mutation and exon skipping in the Fanconi anaemia group C gene. Hum Mol Genet 1993; 2: 797-799
30 Tomiyama Y, Kashi wagi H, Kosugi S, Shiraga M, Kanayama Y, Kurata Y, Matsuzawa Y. Abnormal processing of the glycoprotein lib transcript due to a nonsense mutation in exon 17 associated with Glanzmann’s thrombasthenia. Thromb Haemost 1995; 73: 756-762
31 Naylor JA, Green PM, Rizza CR, Giannelli F. Analysis of factor VIII mRNA reveals defects in every one of 28 haemophilia A patients. Hum Mol Genet 1993; 2: 11-17
32 Dietz HC, Valle D, Francomano CA, Kendzior Jr RJ, Pyeritz RE, Cutting GR. The skipping of constitutive exons in vivo induced by nonsense mutations. Science 1993; 259: 680-683
33 Steingrimsdottir H, Rowley G, Dorago G, Cole J, Lehmann AR. Mutations which alter splicing in the human hypoxanthine-guanine phosphoribosyl-transferase gene. Nucl Acid Res 1992; 20: 1201-1208